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Construction of a rice bacterial artificial chromosome library and identification of clones linked to the Xa‐21 disease resistance locus
Author(s) -
Wang GuoLiang,
Holsten Thomas E.,
Song WenYuan,
Wang HePing,
Ronald Pamela C.
Publication year - 1995
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1046/j.1365-313x.1995.7030525.x
Subject(s) - bacterial artificial chromosome , biology , genetics , locus (genetics) , insert (composites) , transformation (genetics) , telomere , dna , genome , genomic library , library , dna sequencing , chromosome , microbiology and biotechnology , gene , base sequence , engineering , 16s ribosomal rna , mechanical engineering
Summary A bacterial artificial chromosome (BAC) library consisting of 11 000 clones with an average DNA insert size of 125 kb was constructed from rice nuclear DNA. The BAC clones were stable in E. coli after 100 generations of serial growth. Transformation of the BAC clones by electroporation into E. coli was highly efficient and increased with decreasing size of the DNA inserts. The library was evaluated for the presence of organellar, repeated, and telomeric sequences. A very low percentage (<0.3%) of the library consisted of chloroplast and mitochondrial clones. Eighteen BACs were identified that hybridized with an Arabidopsis telomere repeat. Sixteen BACs hybridized with the AA genome‐specific repetitive sequence pOs48 . Twelve clones were isolated that hybridized with three DNA markers linked to the Xa‐21 disease resistance locus. The results indicate that the BAC system can be used to clone and manipulate large pieces of plant DNA efficiently.